Chimeric exchange of coronavirus nsp5 proteases (3CLpro) identifies common and divergent regulatory determinants of protease activity.
Identifieur interne : 002040 ( Main/Exploration ); précédent : 002039; suivant : 002041Chimeric exchange of coronavirus nsp5 proteases (3CLpro) identifies common and divergent regulatory determinants of protease activity.
Auteurs : Christopher C. Stobart ; Nicole R. Sexton ; Havisha Munjal ; Xiaotao Lu ; Katrina L. Molland ; Sakshi Tomar ; Andrew D. Mesecar ; Mark R. DenisonSource :
- Journal of virology [ 1098-5514 ] ; 2013.
Descripteurs français
- KwdFr :
- Alignement de séquences, Animaux, Chimère (), Chimère (génétique), Chimère (métabolisme), Chimère (physiologie), Coronavirus (), Coronavirus (enzymologie), Coronavirus (génétique), Cricetinae, Données de séquences moléculaires, Humains, Infections à coronavirus (médecine vétérinaire), Infections à coronavirus (virologie), Motifs d'acides aminés, Peptide hydrolases (), Peptide hydrolases (génétique), Peptide hydrolases (métabolisme), Phylogénie, Protéines virales (), Protéines virales (génétique), Protéines virales (métabolisme), Souris, Structure tertiaire des protéines, Séquence conservée, Séquence d'acides aminés, Évolution moléculaire.
- MESH :
- enzymologie : Coronavirus.
- génétique : Chimère, Coronavirus, Peptide hydrolases, Protéines virales.
- médecine vétérinaire : Infections à coronavirus.
- métabolisme : Chimère, Peptide hydrolases, Protéines virales.
- physiologie : Chimère.
- virologie : Infections à coronavirus.
- Alignement de séquences, Animaux, Chimère, Coronavirus, Cricetinae, Données de séquences moléculaires, Humains, Motifs d'acides aminés, Peptide hydrolases, Phylogénie, Protéines virales, Souris, Structure tertiaire des protéines, Séquence conservée, Séquence d'acides aminés, Évolution moléculaire.
English descriptors
- KwdEn :
- Amino Acid Motifs, Amino Acid Sequence, Animals, Chimera (classification), Chimera (genetics), Chimera (metabolism), Chimera (physiology), Conserved Sequence, Coronavirus (chemistry), Coronavirus (classification), Coronavirus (enzymology), Coronavirus (genetics), Coronavirus Infections (veterinary), Coronavirus Infections (virology), Cricetinae, Evolution, Molecular, Humans, Mice, Molecular Sequence Data, Peptide Hydrolases (chemistry), Peptide Hydrolases (genetics), Peptide Hydrolases (metabolism), Phylogeny, Protein Structure, Tertiary, Sequence Alignment, Viral Proteins (chemistry), Viral Proteins (genetics), Viral Proteins (metabolism).
- MESH :
- chemical , chemistry : Peptide Hydrolases, Viral Proteins.
- chemistry : Coronavirus.
- classification : Chimera, Coronavirus.
- enzymology : Coronavirus.
- genetics : Chimera, Coronavirus, Peptide Hydrolases, Viral Proteins.
- metabolism : Chimera, Peptide Hydrolases, Viral Proteins.
- physiology : Chimera.
- veterinary : Coronavirus Infections.
- virology : Coronavirus Infections.
- Amino Acid Motifs, Amino Acid Sequence, Animals, Conserved Sequence, Cricetinae, Evolution, Molecular, Humans, Mice, Molecular Sequence Data, Phylogeny, Protein Structure, Tertiary, Sequence Alignment.
Abstract
Human coronaviruses (CoVs) such as severe acute respiratory syndrome CoV (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV) cause epidemics of severe human respiratory disease. A conserved step of CoV replication is the translation and processing of replicase polyproteins containing 16 nonstructural protein domains (nsp's 1 to 16). The CoV nsp5 protease (3CLpro; Mpro) processes nsp's at 11 cleavage sites and is essential for virus replication. CoV nsp5 has a conserved 3-domain structure and catalytic residues. However, the intra- and intermolecular determinants of nsp5 activity and their conservation across divergent CoVs are unknown, in part due to challenges in cultivating many human and zoonotic CoVs. To test for conservation of nsp5 structure-function determinants, we engineered chimeric betacoronavirus murine hepatitis virus (MHV) genomes encoding nsp5 proteases of human and bat alphacoronaviruses and betacoronaviruses. Exchange of nsp5 proteases from HCoV-HKU1 and HCoV-OC43, which share the same genogroup, genogroup 2a, with MHV, allowed for immediate viral recovery with efficient replication albeit with impaired fitness in direct competition with wild-type MHV. Introduction of MHV nsp5 temperature-sensitive mutations into chimeric HKU1 and OC43 nsp5 proteases resulted in clear differences in viability and temperature-sensitive phenotypes compared with MHV nsp5. These data indicate tight genetic linkage and coevolution between nsp5 protease and the genomic background and identify differences in intramolecular networks regulating nsp5 function. Our results also provide evidence that chimeric viruses within coronavirus genogroups can be used to test nsp5 determinants of function and inhibition in common isogenic backgrounds and cell types.
DOI: 10.1128/JVI.02050-13
PubMed: 24027335
Affiliations:
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Le document en format XML
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Sexton</name></author><author><name sortKey="Munjal, Havisha" sort="Munjal, Havisha" uniqKey="Munjal H" first="Havisha" last="Munjal">Havisha Munjal</name></author><author><name sortKey="Lu, Xiaotao" sort="Lu, Xiaotao" uniqKey="Lu X" first="Xiaotao" last="Lu">Xiaotao Lu</name></author><author><name sortKey="Molland, Katrina L" sort="Molland, Katrina L" uniqKey="Molland K" first="Katrina L" last="Molland">Katrina L. Molland</name></author><author><name sortKey="Tomar, Sakshi" sort="Tomar, Sakshi" uniqKey="Tomar S" first="Sakshi" last="Tomar">Sakshi Tomar</name></author><author><name sortKey="Mesecar, Andrew D" sort="Mesecar, Andrew D" uniqKey="Mesecar A" first="Andrew D" last="Mesecar">Andrew D. Mesecar</name></author><author><name sortKey="Denison, Mark R" sort="Denison, Mark R" uniqKey="Denison M" first="Mark R" last="Denison">Mark R. 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(genetics)</term><term>Peptide Hydrolases (metabolism)</term><term>Phylogeny</term><term>Protein Structure, Tertiary</term><term>Sequence Alignment</term><term>Viral Proteins (chemistry)</term><term>Viral Proteins (genetics)</term><term>Viral Proteins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Alignement de séquences</term><term>Animaux</term><term>Chimère ()</term><term>Chimère (génétique)</term><term>Chimère (métabolisme)</term><term>Chimère (physiologie)</term><term>Coronavirus ()</term><term>Coronavirus (enzymologie)</term><term>Coronavirus (génétique)</term><term>Cricetinae</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Infections à coronavirus (médecine vétérinaire)</term><term>Infections à coronavirus (virologie)</term><term>Motifs d'acides aminés</term><term>Peptide hydrolases ()</term><term>Peptide hydrolases (génétique)</term><term>Peptide hydrolases (métabolisme)</term><term>Phylogénie</term><term>Protéines 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aminés</term><term>Évolution moléculaire</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Human coronaviruses (CoVs) such as severe acute respiratory syndrome CoV (SARS-CoV) and Middle East respiratory syndrome CoV (MERS-CoV) cause epidemics of severe human respiratory disease. A conserved step of CoV replication is the translation and processing of replicase polyproteins containing 16 nonstructural protein domains (nsp's 1 to 16). The CoV nsp5 protease (3CLpro; Mpro) processes nsp's at 11 cleavage sites and is essential for virus replication. CoV nsp5 has a conserved 3-domain structure and catalytic residues. However, the intra- and intermolecular determinants of nsp5 activity and their conservation across divergent CoVs are unknown, in part due to challenges in cultivating many human and zoonotic CoVs. To test for conservation of nsp5 structure-function determinants, we engineered chimeric betacoronavirus murine hepatitis virus (MHV) genomes encoding nsp5 proteases of human and bat alphacoronaviruses and betacoronaviruses. Exchange of nsp5 proteases from HCoV-HKU1 and HCoV-OC43, which share the same genogroup, genogroup 2a, with MHV, allowed for immediate viral recovery with efficient replication albeit with impaired fitness in direct competition with wild-type MHV. Introduction of MHV nsp5 temperature-sensitive mutations into chimeric HKU1 and OC43 nsp5 proteases resulted in clear differences in viability and temperature-sensitive phenotypes compared with MHV nsp5. These data indicate tight genetic linkage and coevolution between nsp5 protease and the genomic background and identify differences in intramolecular networks regulating nsp5 function. Our results also provide evidence that chimeric viruses within coronavirus genogroups can be used to test nsp5 determinants of function and inhibition in common isogenic backgrounds and cell types. </div></front></TEI><affiliations><list></list><tree><noCountry><name sortKey="Denison, Mark R" sort="Denison, Mark R" uniqKey="Denison M" first="Mark R" last="Denison">Mark R. Denison</name><name sortKey="Lu, Xiaotao" sort="Lu, Xiaotao" uniqKey="Lu X" first="Xiaotao" last="Lu">Xiaotao Lu</name><name sortKey="Mesecar, Andrew D" sort="Mesecar, Andrew D" uniqKey="Mesecar A" first="Andrew D" last="Mesecar">Andrew D. Mesecar</name><name sortKey="Molland, Katrina L" sort="Molland, Katrina L" uniqKey="Molland K" first="Katrina L" last="Molland">Katrina L. Molland</name><name sortKey="Munjal, Havisha" sort="Munjal, Havisha" uniqKey="Munjal H" first="Havisha" last="Munjal">Havisha Munjal</name><name sortKey="Sexton, Nicole R" sort="Sexton, Nicole R" uniqKey="Sexton N" first="Nicole R" last="Sexton">Nicole R. Sexton</name><name sortKey="Stobart, Christopher C" sort="Stobart, Christopher C" uniqKey="Stobart C" first="Christopher C" last="Stobart">Christopher C. Stobart</name><name sortKey="Tomar, Sakshi" sort="Tomar, Sakshi" uniqKey="Tomar S" first="Sakshi" last="Tomar">Sakshi Tomar</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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